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DSMZ培养基383b组分A基础/DSMZ383b组分A基础
Desulfonauticus Medium Base,DSMZ medium 383b Solution A Base,DSMZ Medium 383b/DSMZ Medium383b/DSMZ 383b/DSMZ383b
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配套试剂
- 1. SD029 - 微量元素溶液SL-10/微量元素溶液SL10 Trace elements solution SL-10/Trace elements solution SL10
- 2. SD052 - 亚硒酸盐-钨酸盐溶液 Selenite-tungstate solution
- 3. SD104 - 10%硫化钠溶液（0.5g/支） 10% Na2S solution
- 4. SD092 - Wolin氏维生素溶液(10x) Wolin's vitamin solution(10x)
产品详情
培养基配方（每升）
Na2SO4 3.0g
KH2PO4 0.2g
NH4Cl 0.3g
NaCl 21.0g
MgCl2·6H2O 3.0g
KCl 0.5g
CaCl2·2H2O 0.15g
Sodium resazurin(刃天青钠)0.0005g
KH2PO4 0.2g
NH4Cl 0.3g
NaCl 21.0g
MgCl2·6H2O 3.0g
KCl 0.5g
CaCl2·2H2O 0.15g
Sodium resazurin(刃天青钠)0.0005g
使用说明
称取本品28.15g于970ml去离子水中，加入微量元素溶液SL-10(SD029)和亚硒酸盐-钨酸盐溶液(SD052)各1支，搅拌溶解，通入80%H2和20%CO2的混合气体不低于30分钟使pH低于6.0，装入厌氧容器中，121℃灭菌15分钟。
制备DSMZ培养基383b时，另制备并加入溶液B(1g碳酸钠、20ml水，121℃灭菌15分钟)、C(1支SD092)以及D(取4mlSD104)，混合，使用无菌的酸碱溶液调节pH至7.0-7.2。备用
制备DSMZ培养基383b时，另制备并加入溶液B(1g碳酸钠、20ml水，121℃灭菌15分钟)、C(1支SD092)以及D(取4mlSD104)，混合，使用无菌的酸碱溶液调节pH至7.0-7.2。备用
特殊说明
383b: DESULFONAUTICUS MEDIUM
1. Solution A is sparged with 80% H2 and 20% CO2 gas mixture to reach a pH below 6 (at
least 30 min), then dispensed under the same gas atmosphere into anoxic Hungate-type
tubes or serum vials to 30% of their volume and autoclaved. Solution B is autoclaved
separately under 80% N2 and 20% CO2 gas atmosphere. Solutions C and E are autoclaved
under 100% N2 gas. Solution D is prepared under 100% N2 gas and filter-sterilized. To
complete the medium appropriate amounts of solutions B to E are added to the sterile
solution A in the sequence as indicated. Final pH of the medium should be 7.0 – 7.2.
2. After inoculation pressurize vials to 2 bar overpressure with sterile 80% H2 and 20% CO2
gas mixture.
3. Note: Addition of 10 - 20 mg sodium dithionite per liter (e.g. from 5% (w/v) solution
freshly prepared under N2 and filter-sterilized) may stimulate growth of some strains at the
beginning. For transfers use 5 - 10% inoculum. Incubate all strains in the dark.
For DSM 4206, DSM 15269: Supplement medium with 1.00 g/l Na-acetate, 2.00 g/l
Trypticase peptone and 2.00 g/l yeast extract added to the autoclaved medium from
sterile anoxic stock solutions prepared under N2 gas.
For DSM 15286, DSM 21156, DSM 101864: Supplement medium with 1.00 g/l Na-acetate
and 1.00 ml/l seven vitamins solution (see medium 503) added to the autoclaved medium
from sterile and anoxic stock solutions. Vitamins are sterilized by filtration. Adjust pH of
medium to 6.5 - 6.7 prior to inoculation.
For DSM 111414: Adjust pH of final medium to 6.6.
Solution A
Solution B
Solution C
Wolin's vitamin solution (10x) 1.00 ml
Solution D
Selenite-tungstate solution (from medium 385)
Trace element solution SL-10 (from medium 320)
First dissolve FeCl2 in the HCl, then dilute in water, add and dissolve the other salts. Finally
make up to 1000.00 ml.
Wolin's vitamin solution (10x) (from medium 120)
p-Aminobenzoic acid 50.00 mg
(DL)-alpha-Lipoic acid 50.00 mg
Distilled water 1000.00 ml
|
Solution A Solution B Solution C Solution D |
972.00 20.00 1.00 13.00 |
ml ml ml ml |
least 30 min), then dispensed under the same gas atmosphere into anoxic Hungate-type
tubes or serum vials to 30% of their volume and autoclaved. Solution B is autoclaved
separately under 80% N2 and 20% CO2 gas atmosphere. Solutions C and E are autoclaved
under 100% N2 gas. Solution D is prepared under 100% N2 gas and filter-sterilized. To
complete the medium appropriate amounts of solutions B to E are added to the sterile
solution A in the sequence as indicated. Final pH of the medium should be 7.0 – 7.2.
2. After inoculation pressurize vials to 2 bar overpressure with sterile 80% H2 and 20% CO2
gas mixture.
3. Note: Addition of 10 - 20 mg sodium dithionite per liter (e.g. from 5% (w/v) solution
freshly prepared under N2 and filter-sterilized) may stimulate growth of some strains at the
beginning. For transfers use 5 - 10% inoculum. Incubate all strains in the dark.
For DSM 4206, DSM 15269: Supplement medium with 1.00 g/l Na-acetate, 2.00 g/l
Trypticase peptone and 2.00 g/l yeast extract added to the autoclaved medium from
sterile anoxic stock solutions prepared under N2 gas.
For DSM 15286, DSM 21156, DSM 101864: Supplement medium with 1.00 g/l Na-acetate
and 1.00 ml/l seven vitamins solution (see medium 503) added to the autoclaved medium
from sterile and anoxic stock solutions. Vitamins are sterilized by filtration. Adjust pH of
medium to 6.5 - 6.7 prior to inoculation.
For DSM 111414: Adjust pH of final medium to 6.6.
Solution A
|
Na2SO4 KH2PO4 NH4Cl NaCl MgCl2 x 6 H2O KCl CaCl2 x 2 H2O Trace element solution SL-10 Selenite-tungstate solution Sodium resazurin (0.1% w/v) Distilled water |
3.00 0.20 0.30 21.00 3.00 0.50 0.15 1.00 1.00 0.50 970.00 |
g g g g g g g ml ml ml ml |
Solution B
|
Na2CO3 Distilled water |
1.00 20.00 |
g ml |
Wolin's vitamin solution (10x) 1.00 ml
Solution D
|
Na2S x 9 H2O Distilled water |
0.40 13.00 |
g ml |
|
NaOH Na2SeO3 x 5 H2O Na2WO4 x 2 H2O Distilled water |
0.50 3.00 4.00 1000.00 |
g mg mg ml |
|
HCl (25%) FeCl2 x 4 H2O ZnCl2 MnCl2 x 4 H2O H3BO3 CoCl2 x 6 H2O CuCl2 x 2 H2O NiCl2 x 6 H2O Na2MoO4 x 2 H2O Distilled water |
10.00 1.50 70.00 100.00 6.00 190.00 2.00 24.00 36.00 990.00 |
ml g mg mg mg mg mg mg mg ml |
make up to 1000.00 ml.
Wolin's vitamin solution (10x) (from medium 120)
|
Biotin Folic acid Pyridoxine hydrochloride Thiamine HCl Riboflavin Nicotinic acid Calcium D-(+)-pantothenate Vitamin B12 |
20.00 20.00 100.00 50.00 50.00 50.00 50.00 1.00 |
mg mg mg mg mg mg mg mg |
(DL)-alpha-Lipoic acid 50.00 mg
Distilled water 1000.00 ml
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